The present invention relates to a novel herbal composition for the treatment of cancer. The present invention particularly relates to an herbal formulation comprising mixture of lignans isolated from the plant Cedrus Deodara. 
Cancer or neoplasm is the malignant new growth anywhere and elsewhere in the body system. It is characterized by unregulated proliferation of cells and a growing public health problem whose estimated worldwide new incidence is about six million cases per year. In most of the countries, cancer is second only to heart disease as cause of death. It can arise in any organ of the body but some sites are prone to attack than others such as breast, throat, intestine, leukocytes etc. Each cancer is propagated from a single cell that cut at some stage, it becomes free from its territorial restraints, which form a family of cells that multiply without limits and appear in the form of tumors.
During the transition from normal cell to a tumor cell a profound and heritable change occurs which allows a tumor cell to determine its own activities largely irrespective of the laws that govern so precisely the growth of all normal cells in an organism. This newly acquired property, which is known as autonomy, is the most important single characteristic of tumor cells since without it there would be no tumors. Another distinguishing characteristic of tumor cells is their lack of perfect form of function. The differences that exist between cancer and normal cells are that, compared to normal cells, cancer cells have a) low pH b) greater free radical character c) tumor produced hormone peptides d) tumor associated antigens e) lower calcium ion and higher potassium ion concentration f) different potassium isotope ratios g) elevated amounts of methylated nucleotides h) higher concentrations of plasma microproteins and mucopolysacharides i) greater need of exogenous zinc and j) high biowater content.
Many of the gross causes of cancer, such as dietary, environmental, occupational exposure to certain chemical substances or forms of electromagnetic radiation, have been elucidated through epidemiological studies. It is imperative, therefore, that they be identified and eliminated from the environment in so far as that is possible in modern industrial societies. In the annals of therapy, a quest to conquer, the impasse of cancer has been always fascinated by and large all disciplines of scientific community, especially natural product chemists. In 19th and 20th century, lot of research work has been carried out to find out the driving force behind this dreadful disease as well as large number of drugs have been introduced to counter its menace.
It is worthwhile at this juncture to look briefly at a few most powerful chemotherapeutic agents, which have been of paramount importance to the mankind and also to the researchers who have been actively involved in the synthesis and isolation of anticancer drugs. Lignans have been isolated from a number of plants (Achenbach, H., Waibel, R. and Meusah, I. Phytochemistry, 22 (3), 749-753 (1983); Nishibe, S., Hisada, S. and Inagaki, I. Phytochemistry, 10, 2231-2232 (1971); Barrero, A. F., Haidour, A. and Dorado, M. M. J. Nat. Prod. 42,159-162(1979)).
Recent studies on the biological activities of lignans prove beyond doubt the efficacy of these phytochemicals as cytotoxic agents (Macrae, W. D. and Towers, G. H. N. Phytochemistry 23 (6), 1207-1220 (1984)). Also recently lignans have been isolated from human urine and blood. This fact also suggests that lignans are playing a definite role in human physiology.
With the above background, the applicants have focussed their attention towards the identification and isolation of potent cytotoxic agents from plants. Literature data suggests the presence secoisolariciresinol in Cedrus deodara (Agarwal, P. K. and Rastogi, R. P. Phytochemistry, 21 (6), 1459-1461 (1982)) which is in all likelihood may be an artefact, is a proven antioxidant. Further investigation on Cedrus Deodara carried out by us led us to the isolation of a new lignan mixture comprising essentially (xe2x88x92)-Matairesinol, (xe2x88x92)-Wikshtronol and Dibenzyl butyrolactol in an extremely significant yield, Cedrus deodara is also a new source for these lignans.
(xe2x88x92)-Wikstromol was first reported from wikstroemia viridiflora (Nishibe, S., Hisada, S. and Inagaki, I. Phytochemistry, 10, 2231-2232 (1971)). Matairesinol was isolated from a number of sources before (Nishibe, S., Hisada, S. and Inagaki, I. Phytochemistry, 10, 2231-2232 (1971); Tandon, S. and Rastogi, R. P. Phytochemistry, 15, 1789-1791 (1976)). The isolation of the dibenzylbutyrolactol [4,4xe2x80x2,9-trihydroxy-3,3xe2x80x2-dimethoxy-9,9xe2x80x2-epoxy lignan] was reported only once previously from the wood of Abies pinsapo (Barrero, A. F., Haidour, A. and Dorado, M. M. J. Nat. Prod. 42, 159-162 (1979)).
Keeping in mind the high yields of the lignans from Cedrus deodara and also the excellent biological activities and lignans in general (Macrae, W. D. and Towers, G. H. N. Phytochemistry 23(6), 1207-1220 (1984).
Main object of the invention relates to herbal compositions for the treatment of human cancer cells.
Another object of the invention relates to herbal compositions for the treatment of human cancer cells obtained from a plant source.
Another object of the invention relates to in-vitro cytotoxicity of lignan mixtures isolated from Cedrus deodara against various human cancer cell lines.
Still another object of the invention relates to cytotoxicity of individual lignans isolated from Cedrus deodara against various human cancer cell lines.
Still another embodiment of the invention relates to a method of isolation of the active lignan mixture from the plant source namely Cedrus deodara. 
Yet another object of the invention relates to a synergistic lignan composition obtained from plant Cedrus deodara for inhibiting the growth of human cancer cells.
Yet another object of the invention relates to a composition comprising individual lignans isolated from plant Cedrus deodara for inhibiting the growth of human cancer cells.
Yet another embodiment relates to method of treating mammals, particularly human beings affected by cancer.
Accordingly, the present invention provides a novel synergistic composition of lignans exhibiting anticancer activities for breast, cervix, neuroblastoma, colon, liver, lung, mouth, ovary and prostate cancer obtained from the plant extract of Cedrus deodra, said composition comprising_of
(a) (xe2x88x92) -Matairesinol in the range of 9 to 13% by weight,
(b) (xe2x88x92) -Wikstromol in the range of 75 to 79% by weight,
(c) Dibenzylbutyrolactol in the range of 7 to 11% by weight, and
(d) Unidentified material in the range of 2.6 to 3% by weight,
Further, the synergistic composition of lignan may be used in combination with pharmaceutically acceptable carriers for inhibiting various human cancer cell lines.
An attempt is made to establish anticancer activity to the lignan mixture isolated from Cedrus deodara. The individual constituents isolated from the chloroform extract of Cedrus deodara wood have shown lesser level of activity compared to the lignan mixture establishing the principle of synergy.
Accordingly, the present invention provides a novel synergistic composition of lignans exhibiting anticancer activities for breast, cervix, neuroblastoma, colon, liver, lung, mouth, ovary and prostate cancer obtained from the plant extract of Cedrus deodra, said composition comprising_of
(e) (xe2x88x92) -Matairesinol in the range of 9 to 13% by weight,
(f) (xe2x88x92) -Wikstromol in the range of 75 to 79% by weight,
(g) Dibenzylbutyrolactol in the range of 7 to 11% by weight, and
(h) Unidentified material in the range of 2.6 to 3% by weight,
In another embodiment, wherein the synergistic lignans composition inhibits the growth of cancer cells of breast up to 80% at a concentration ranging from 30-100 xcexcg/ml.
Still another embodiment, the breast cell line is selected from group consisting of MCF-7 and T-47-D.
Yet another embodiment, wherein the synergistic lignans composition inhibits the growth of cancer cells of cervix up to 89% at a concentration ranging from 30-100 xcexcg/ml.
Yet another embodiment, the cervix cell line is selected from the group consisting of Hela and SiHa.
Yet another embodiment, wherein the synergistic lignans composition inhibits the growth of cancer cells of neuroblastoma up to 96% at a concentration ranging from 30-100 xcexcg/ml.
Yet another embodiment, wherein cancer cell line of neuroblastoma is selected from the group consisting of SF-539, SKNMC, IMR-32, SKNSH and SNB-78.
Yet another embodiment, wherein the synergistic lignan composition inhibits the growth of cancer cells of colon up to 97% at a concentration ranging from 30-100 xcexcg/ml.
Yet another embodiment, wherein cancer cell line of colon is selected from the group consisting of Colo-205, HCT-15, HT-29 and SW-620.
Yet another embodiment, wherein the synergistic lignan composition inhibits the growth of cancer cells of liver up to 73% at a concentration ranging from 30-100 xcexcg/ml.
Yet another embodiment, wherein the cancer cell line of liver is selected from the group consisting of Hep-2 and Hep-G-2.
Yet another embodiment, wherein the synergistic lignan composition inhibits the growth of cancer cells of lung up to 83% at a concentration ranging from 30-100 xcexcg/ml.
Yet another embodiment, wherein the cancer cell line of lung is selected from the group consisting of A-549, NCI-H23 and HOP-18.
Yet another embodiment, wherein the synergistic lignan composition inhibits the growth of cancer cells of mouth up to 100% at a concentration ranging from 30-100 xcexcg/ml.
Yet another embodiment, wherein the cancer cell line of mouth is KB.
Yet another embodiment, the synergistic lignan composition inhibits the growth of cancer cells of ovary up to 96% at a concentration ranging from 30-100 xcexcg/ml.
Yet another embodiment, wherein the cancer cell line of ovary is selected from the group consisting of OVCAR-5, NIH-OVCAR-3 and SK-OV-3.
Yet another embodiment, wherein the synergistic lignan composition inhibits the growth of cancer cells of prostrate up to 98% at a concentration of ranging from 30-100 xcexcg/ml.
Yet another embodiment, wherein the cancer cells line of prostrate tissue is selected from the group consisting of DU-145 and PC-3.
Yet another embodiment, wherein the synergistic composition of lignan is administered to the patient in combination with a pharmaceutically acceptable additive, carrier, diluent, solvent, filter, lubricant, excipient, binder, or stabilizer.
Yet another embodiment, wherein synergistic lignan composition can be administered systemically and/or orally or any other suitable method.
Yet another embodiment, wherein the subjects are selected from animals or mammals preferably humans.
One more embodiment of the invention relates to a composition exhibiting anti cancer activities for cancer cell lines selected mainly from the group consisting breast, cervix, neuroblastoma, colon, liver and lung, said composition comprising pharmaceutically effective dosage of (xe2x88x92)-wikstromol or a formulation containing.(xe2x88x92)-wikstromol.
Another embodiment of the invention, wherein the cancer cell line used are selected from group consisting breast cells MCF-7 and ZR-75-1; cervix cell SiHa; neuroblastoma cells SKNMC and IMR-32; colon cells colo-205HF-29 and SW-620; liver cell Hep-2; and lung cell A-549.
Still another embodiment of the invention, wherein above said composition inhibits the growth of cancer cells of breast up to 51% at a concentration of about 100 xcexcg/ml.
Yet another embodiment of the invention, wherein said composition inhibits the growth of cancer cells of cervix up to 37% at a concentration of about 100 xcexcg/ml.
Yet another embodiment of the invention, wherein said composition inhibits the growth of cancer cells of neuroblastoma up to 56% at a concentration of about 100 xcexcg/ml.
Yet another embodiment of the invention, wherein said composition inhibits the growth of cancer cells of colon up to 67% at a concentration of about 100 xcexcg/ml.
Yet another embodiment of the invention, wherein said composition inhibits the growth of cancer cells of liver up to 46% at a concentration of about 100 xcexcg/ml.
Yet another embodiment of the invention, wherein said composition inhibits the growth of cancer cells of lung up to 56% at a concentration of about 100 xcexcg/ml.
Yet another embodiment of the invention, wherein said composition inhibits is administered in combination with a pharmaceutically acceptable carriers, additives, diluents, solvents, filters, lubricants, excipients, binders and stabilizers.
Yet another embodiment, said composition can be administered systematically and/or orally or any other suitable method.
Yet another embodiment, wherein the subjects are selected from animals or mammals preferably humans.
One more embodiment of the present invention provides a composition exhibiting anti cancer activities for cancer cell lines selected mainly from the group consisting breast, cervix, neuroblastoma, colon, liver and lung, said composition comprising a pharmaceutically effective dosage of (xe2x88x92)- Matairesinol or a formulation containing (xe2x88x92)- Matairesinol.
Yet another embodiment, wherein the cancer cell lines used are selected from the group comprising of breast cells MCF-7 and ZR-75-1; cervix cell SiHa; neuroblastoma cells SKNMC and IMR-32; colon cells Colo-205, HT-29 and SW-620; liver cell Hep-2; and lung cell A-549.
Yet another embodiment, wherein composition containing (xe2x88x92)- Matairesinol inhibits the growth of cancer cells of breast up to 62% at a concentration of about 100 xcexcg/ml
Yet another embodiment, wherein above said composition inhibits the growth of cancer cells of cervix up to 63% at a concentration of about 100 xcexcg/ml
Yet another embodiment, wherein above said composition inhibits the growth of cancer cells of neuroblastoma up to 77% at a concentration of about 100 xcexcg/ml
Yet another embodiment, wherein above said composition inhibits the growth of cancer cells of colon up to 93% at a concentration of about 100 xcexcg/ml
Yet another embodiment, wherein above said composition inhibits the growth of cancer cells of liver up to 64% at a concentration of about 100 xcexcg/ml
Yet another embodiment, wherein above said composition inhibits the growth of cancer cells of lung up to 65% at a concentration of about 100 xcexcg/ml
Yet another embodiment, wherein above said composition is administered to the patient in combination with a pharmaceutically acceptable carriers additives, diluents, solvents, filters, lubricants, excipients, binders and stabilizers.
Yet another embodiment, wherein above said composition may be administered systematically or orally.
Yet another embodiment, wherein the subjects are selected from animals or mammals preferably humans
One more embodiment of the invention relates to a process for isolation of synergistic lignan composition from the plant extract of Cedrus deodra comprises steps of:
(a) powdering the plant Cedrus deodra, 
(b) extracting the powdered plant in a soxhlet apparatus successively with hydrocarbon solvent followed by halogenated solvent,
(c) concentrating the extract of hydrocarbon solvent and halgenated solvent separately, and
(d) purifying halogenated solvent extract by chromatography on an adsorbent by eluting with mixture of organic solvents to yield (xe2x88x92)- Matairesinol 1 to 2% by weight, (xe2x88x92)- Wikstromol 10 to 14% by weight, Dibenzylbutyrolactol 1 to 2% by weight and unidentified material 0.2 to 0.3% by weight with respect to extract and which together constitute as synergistic lignan composition.
Another embodiment of the invention, in step (a) wherein the plant part is wood and in step (b) the hydrocarbon solvent is selected from group of hexane and petroleum ether, preferably hexane.
Still another embodiment of the invention, the halogenated solvent is selected from carbon tetrachloride, methylene chloride or chloroform, preferably chloroform.
Still another embodiment of the invention, in step (c) the extracts are concentrated under vacuum and the purification is carried out using silica gel as an adsorbent and eluting with different proportions of chloroform-methanol mixture to get pure compounds (.)wikstromol, (xe2x88x92)-metairesinol, dibenzylbutyrolactol, an unidentified material and which together constitute as synergistic lignan composition.
One more embodiment of the invention provide a method of treating a patient with cancer mainly breast, cervix, neuroblastoma, colon, liver, lung, mouth, ovary and prostate, said method comprising administering a pharmaceutically effective dosage of synergistic composition of lignan composition to the patient.
In another embodiment of the invention, wherein the said synergistic lignan composition is used in combination with pharmaceutically acceptable carriers.
Still another embodiment of the invention, wherein synergistic lignan composition may be administered systemically and/or orally or any other suitable method.
Still another embodiment of the invention, wherein the subjects are selected from animals or mammals preferably humans.
Yet another embodiment of the invention, wherein said synergistic lignan composition inhibits the growth of cancel cells of breast up to 80% at a concentration ranging from 30-100 xcexcg/ml.
Yet another embodiment of the invention, breast cell lines inhibited are MCF-7 and T-47-D.
Yet another embodiment of the invention, wherein synergistic lignan composition inhibits the growth of cancel cells of cervix up to 81% at a concentration ranging from 30-100 xcexcg/ml.
Yet another embodiment of the invention, cervix cell lines inhibited are Hela and SiHa.
Yet another embodiment of the invention, wherein synergistic lignan composition inhibits the growth of cancel cells of neuroblastoma up to 92% at a concentration ranging from 30-100 xcexcg/ml.
Yet another embodiment of the invention, wherein neuroblastoma cell lines inhibited are SF-539, SKNMC, IMR-32, SKNSH and SNB-78.
Yet another embodiment of the invention, wherein synergistic lignan composition inhibits the growth of cancel cells of colon up to 95% at a concentration ranging from 30-100 xcexcg/ml.
Yet another embodiments of the invention, wherein colon cell lines inhibited are Colo-205, HCT-15, HT-29 and SW-620.
Yet another embodiment of the invention, wherein synergistic lignan composition inhibits the growth of cancel cells of liver up to 73% at a concentration ranging from 30-100 xcexcg/ml.
Yet another embodiment of the invention, wherein liver cell line inhibited is Hep-2.
Yet another embodiment of the invention, wherein synergistic lignan composition inhibits the growth of cancel cells of lung up to 92% at a concentration ranging from 30-100 xcexcg/ml.
Yet another embodiment of the invention, wherein lung cell lines inhibited are A-549 and NCI-H23.
Yet another embodiment of the invention, wherein synergistic lignan composition inhibits the growth of cancel cells of mouth up to 99% at a concentration ranging from 30-100 xcexcg/ml.
Yet another embodiment of the invention, wherein mouth cell line inhibited is KB
Yet another embodiment of the invention, wherein synergistic lignan composition inhibits the growth of cancel cells of ovary up to 96% at a concentration ranging from 30-100 xcexcg/ml.
Yet another embodiment of the invention, wherein ovary cell lines inhibited are OVCAR5 and SK-OV-3.
Yet another embodiment of the invention, wherein synergistic lignan composition inhibits the growth of cancel cells of prostrate up to 98% at a concentration ranging from 30-100 xcexcg/ml
Yet another embodiment of the invention, wherein prostrate cell lines inhibited are DU-145 and PC-3.
Yet another embodiment of the invention, wherein synergistic lignan composition is administered to the patient in combination with a pharmaceutically acceptable additive, carrier, diluent, solvent, filter, lubricant, excipient, binder, or stabilizer.
Yet another embodiment of the invention, wherein amount of synergistic composition administered is ranging from 10 to 500 mg/kg body weight for at least one dose per day.
Yet another embodiment of the invention, wherein, amount of synergistic composition administered is preferably ranging from 50 to 350 mg/kg body weight.
Another more embodiment of the invention relates to a method of treating a patient with cancer mainly breast, cervix, neuroblastoma, colon, liver, lung, mouth, ovary and prostate, said method comprising administering a pharmaceutically effective dosage of (xe2x88x92)-wikstromol or a composition containing (xe2x88x92)-woikstromol. to the patient.
Still another embodiment, wherein the said composition inhibits the growth of cancer cells consisting of breast cells, cervix cells, neuroblastoma cells, colon cells, liver cell and lung cell up to 51%, 37%, 56%, 67%,46% and 56% respectively.
Still another embodiment, wherein the cancer cell lines are selected from the group comprising of breast cells MCF-7 and ZR-75-1; cervix cell SiHa; neuroblastoma cells SKNMC and IMR-32; colon cells Colo-205, HT-29 and SW-620; liver cell Hep-2; and lung cell A-549.
Yet another embodiment, wherein the composition is used singly or in combination with pharmaceutically acceptable carriers.
Yet another embodiment, wherein the composition can be administered systematically and/or orally or any other suitable method.
Yet another embodiment, wherein the subjects are selected from animals or mammals preferably humans.
Yet another embodiment, wherein the amount of composition administered is ranging from 10 to 500 mg/kg body weight at least once in a day.
Yet another embodiment, wherein the amount of composition administered is preferably ranging from 75 to 300 mg/kg body weight at least once in a day.
Yet another embodiment, wherein the (xe2x88x92)-wikstromol is administered to the patient in combination with a pharmaceutically acceptable carriers additives, diluents, solvents, filters, lubricants, excipients, binders and stabilizers.
One more embodiment relates to a method of treating a patient with cancer mainly breast, cervix, neuroblastoma, colon, liver and lung, said method comprising administering a pharmaceutically effective dosage of (xe2x88x92)-Matairesinol or a composition containing (xe2x88x92)-Matairesinol to the patient.
Another embodiment of the invention, wherein the said composition inhibits the growth of cancer cells consisting of breast cells, cervix cells, neuroblastoma cells, colon cells, liver cell and lung cell up to 62%, 63%, 77%, 93%, 64% and 65% respectively.
Yet another embodiment, wherein the cancer cell lines are selected from the group comprising of breast cells MCF-7 and ZR-75-1; cervix cell SiHa; neuroblastoma cells SKNMC and IMR-32; colon cells Colo-205, HT-29 and SW-620; liver cell Hep-2; and lung cell A-549.
Yet another embodiment, the said composition is used singly or in combination with pharmaceutically acceptable carriers.
Yet another embodiment the said composition may be administered systematically or orally.
Yet another embodiment, wherein the subjects are selected from animals or mammals preferably humans.
Yet another embodiment, the amount of composition administered is ranging from 10 to 500 mg/kg body weight at least once in a day.
Yet another embodiment, the amount of composition administered is preferably ranging from 75 to 300 mg/kg body weight at least once in a day.
Yet another embodiment, the said composition is administered to the patient in combination with a pharmaceutically acceptable carriers additives, diluents, solvents, filters, lubricants, excipients, binders and stabilisers.
The present invention embodies isolation of new cytotoxic mixture from an entirely new source. The lignan mixture, in vitro, significantly inhibited the growth of number of human cancer cell lines (breast: MCF-7 and T-47-D, cervix: Hela and SiHa, neuroblastoma: SF-539, SKNMC, IMR-32, SKNSH and SNB-78, colon: Colo-205, HCT-15, HT-29 and SW-620, liver: Hep-2, lung: A-549 and NCI-H23, mouth: KB, ovary: OVCAR5 and SK-OV-3 and prostate: DU-145 and PC-3) representing different organs.
The present invention relates to a synergistic composition comprising (xe2x88x92)-Matairesinol, (xe2x88x92)-Wikstromol, Dibenzylbutyrolactol and unidentified material providing a unexpected results of showing enhanced cytotoxicity against cancer cell lines, which is substantiated by remarkable cytotoxicity results against cancer cell lines selected from Breast, Cervix, Neuroblastoma, Colon, Liver, Lung, Oral, Ovary and Prostate tissues. In fact, the compositions is synergistic because the activity is remarkable and such surprisingly enhanced activity of the composition cannot be expected from the mere aggregation of the properties of the individual ingredients. In other words, the composition does not possess the mere addition of the properties of its ingredients, but an enhanced activity, which further substantiates the efficacy of the synergistic composition isolated. Further, the amounts of the ingredients also impart/contribute for the enhanced activity of the composition.
The following examples are given by way of illustration and therefore should not be construed to limit the scope of the present invention.